Fig. 4 | Communications Biology

Fig. 4

From: Rapid and gentle hydrogel encapsulation of living organisms enables long-term microscopy over multiple hours

Fig. 4

Long-term volumetric imaging of optogenetically stimulated neurons in living C. elegans. Animals expressing Chrimson and GCaMP2.2b in AWA sensory neurons were stimulated by red light and imaged using a diSPIM light-sheet microscope. a Schematic of diSPIM objectives, hydrogel, and red light exposure. b Three-dimensional volumetric view of AWA neurons. ce Maximum intensity projections highlighting ROIs of AWAL and AWR cell bodies and AWAR dendrite, at t = 0 h (c), 6 h (d), and 13.5 h (e) time points. Scale bars, 20 μm. fh Time-lapse recordings of GCaMP signal in each ROI beginning at each time point. The stimulation LED was pulsed for 10 s, once per minute. Continuous recordings lasted for 1 h per time point; here, 30 min are shown for clarity. ik Mean fluorescence (ΔF/F0) is calculated for the 30 stimulation pulses indicated in fh, with shading indicating s.e.m., and red bars above indicating 10-s red light exposure

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